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Fig. 5. Analysis of candidates for downregulation (Odz4 and Epha3) or upregulation (Gdf10) by Hoxd gene products. WISH staining of E12.5 right forelimbs are shown for each gene. Lateral views of genitals (at right in each panel) are at E13.5 for Odz4 and Epha3, and E12.5 for Gdf10. Real-time RT-PCR quantification for each gene is shown as graphs. (A) The Odz4 forelimb expression domain extends more distally in HoxD^Del1-13 mutants at E12.5, and expression is gained in the mutant genitalia. (B) Epha3 is expressed at a higher level in mutant forelimbs. The limb expression domain increases posteriorly (right) and distally, when compared with the wild type (left). Real-time RT-PCR measurements also indicate an increase in genital expression in the HoxD^Del1-13 mutant (^**P<0.01 at E13.5). By WISH, this increase is visualized as a slight broadening of the expression domain in the mutant genitals. (C) Gdf10 expression is weaker in mutant forelimbs and genital buds. Gdf10 is expressed in a broad domain in both wild-type and mutant limbs and genitals, making visualization of differences by WISH difficult, although some decrease in mutant tissues is apparent. Real-time RT-PCR quantification shows highly significant differences (^**P<0.01) in both tissues.

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